Direct and site-specific quantification of RNA 2′-O-methylation by PCR with an engineered DNA polymerase

Aschenbrenner, Joos; Marx, Andreas

Direct and site-specific quantification of RNA 2′-O-methylation by PCR with an engineered DNA polymerase

Files

Aschenbrenner_0-343405.pdf(2.02 MB)

Date

2016

EU project number

339834

Project

EvoEPIGEN - Evolved Replication Systems of Epigenetics

Collections

Chemie

Publication type

Journal article

Publication status

Published

Published in

Nucleic Acids Research ; 44 (2016), 8. - pp. 3495-3502. - ISSN 0301-5610. - eISSN 1362-4962

Abstract

Methylation of the 2'-hydroxyl-group of ribonucleotides is found in all major classes of RNA in eukaryotes and is one of the most abundant posttranscriptional modifications of stable RNAs. In spite of intense studies, the multiple functions of RNA 2'-O-methylation are still not understood. One major obstacle in the field are the technical demanding detection methods, which are typically laborious and do not always deliver unambiguous results. We present a thermostable KlenTaq DNA polymerase variant with significant reverse transcription activity that is able to discriminate 2'-O-methylated from unmethylated RNAs. The engineered enzyme catalyzes DNA synthesis from DNA as well as RNA templates and enables expeditious quantification of 2'-O-methylation of individual nucleotides directly from total RNA extracts by a simple qRT-PCR.

Subject (DDC)

540 Chemistry

Cite This

ISO 690

ASCHENBRENNER, Joos, Andreas MARX, 2016. Direct and site-specific quantification of RNA 2′-O-methylation by PCR with an engineered DNA polymerase. In: Nucleic Acids Research. 44(8), pp. 3495-3502. ISSN 0301-5610. eISSN 1362-4962. Available under: doi: 10.1093/nar/gkw200

BibTex

@article{Aschenbrenner2016-05-05Direc-34853,
  year={2016},
  doi={10.1093/nar/gkw200},
  title={Direct and site-specific quantification of RNA 2′-O-methylation by PCR with an engineered DNA polymerase},
  number={8},
  volume={44},
  issn={0301-5610},
  journal={Nucleic Acids Research},
  pages={3495--3502},
  author={Aschenbrenner, Joos and Marx, Andreas}
}

RDF

<rdf:RDF
    xmlns:dcterms="http://purl.org/dc/terms/"
    xmlns:dc="http://purl.org/dc/elements/1.1/"
    xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#"
    xmlns:bibo="http://purl.org/ontology/bibo/"
    xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#"
    xmlns:foaf="http://xmlns.com/foaf/0.1/"
    xmlns:void="http://rdfs.org/ns/void#"
    xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > 
  <rdf:Description rdf:about="https://kops.uni-konstanz.de/server/rdf/resource/123456789/34853">
    <dc:language>eng</dc:language>
    <dc:creator>Aschenbrenner, Joos</dc:creator>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dc:contributor>Marx, Andreas</dc:contributor>
    <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by-nc/4.0/"/>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/34853/3/Aschenbrenner_0-343405.pdf"/>
    <dc:contributor>Aschenbrenner, Joos</dc:contributor>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2016-07-21T13:10:21Z</dcterms:available>
    <dc:creator>Marx, Andreas</dc:creator>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2016-07-21T13:10:21Z</dc:date>
    <dc:rights>Attribution-NonCommercial 4.0 International</dc:rights>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dcterms:abstract xml:lang="eng">Methylation of the 2'-hydroxyl-group of ribonucleotides is found in all major classes of RNA in eukaryotes and is one of the most abundant posttranscriptional modifications of stable RNAs. In spite of intense studies, the multiple functions of RNA 2'-O-methylation are still not understood. One major obstacle in the field are the technical demanding detection methods, which are typically laborious and do not always deliver unambiguous results. We present a thermostable KlenTaq DNA polymerase variant with significant reverse transcription activity that is able to discriminate 2'-O-methylated from unmethylated RNAs. The engineered enzyme catalyzes DNA synthesis from DNA as well as RNA templates and enables expeditious quantification of 2'-O-methylation of individual nucleotides directly from total RNA extracts by a simple qRT-PCR.</dcterms:abstract>
    <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/34853"/>
    <dcterms:issued>2016-05-05</dcterms:issued>
    <dcterms:title>Direct and site-specific quantification of RNA 2′-O-methylation by PCR with an engineered DNA polymerase</dcterms:title>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/34853/3/Aschenbrenner_0-343405.pdf"/>
  </rdf:Description>
</rdf:RDF>

Bibliography of Konstanz

Yes